Hardness Measurement of Bulk Wheat by Single‐Kernel Visible and Near‐Infrared Reflectance Spectroscopy

Reflectance spectra (400 to 1700 nm) of single wheat kernels collected using the Single Kernel Characterization System (SKCS) 4170 were analyzed for wheat grain hardness using partial least squares (PLS) regression. The wavelengths (650 to 700, 1100, 1200, 1380, 1450, and 1670 nm) that contributed most to the ability of the model to predict hardness were related to protein, starch, and color differences. Slightly better prediction results were observed when the 550–1690 nm region was used compared with 950–1690 nm region across all sample sizes. For the 30‐kernel mass‐averaged model, the hardness prediction for 550–1690 nm spectra resulted in a coefficient of determination (R²) = 0.91, standard error of cross validation (SECV) = 7.70, and relative predictive determinant (RPD) = 3.3, while the 950–1690 nm had R² = 0.88, SECV = 8.67, and RPD = 2.9. Average hardness of hard and soft wheat validation samples based on mass‐averaged spectra of 30 kernels was predicted and compared with the SKCS 4100 reference method (R² = 0.88). Compared with the reference SKCS hardness classification, the 30‐kernel (550–1690 nm) prediction model correctly differentiated (97%) between hard and soft wheat. Monte Carlo simulation technique coupled with the SKCS 4100 hardness classification logic was used for classifying mixed wheat samples. Compared with the reference, the prediction model correctly classified mixed samples with 72–100% accuracy. Results confirmed the potential of using visible and near‐infrared reflectance spectroscopy of whole single kernels of wheat as a rapid and nondestructive measurement of bulk wheat grain hardness.     

Detecting and Segregating Black Tip‐Damaged Wheat Kernels Using Visible and Near‐Infrared Spectroscopy

Detection of individual wheat kernels with black tip symptom (BTS) and black tip damage (BTD) was demonstrated with near‐infrared reflectance spectroscopy (NIRS) and silicon light‐emitting‐diode (LED) based instruments. The two instruments tested, a single‐kernel NIRS instrument (SKNIRS) and a silicon LED‐based single‐kernel high‐speed sorter (SiLED‐SKS) were both developed by the Stored Product Insect and Engineering Research Unit, Center for Grain and Animal Health Research, USDA Agricultural Research Service. BTD was classified into four levels for the study ranging from sound, symptomatic (BTS) at two levels, and damaged (BTD). Discriminant analysis models for the SKNIRS instrument could distinguish sound undamaged kernels well, correctly classifying kernels 80% of the time. Damaged kernels were classified with 67% accuracy and symptomatic kernels at about 44%. Higher classification accuracy (81–87%) was obtained by creating only two groupings: 1) combined sound and lightly symptomatic kernels and 2) combined heavily symptomatic and damaged kernels. A linear regression model was developed from the SiLED‐SKS sorted fractions to predict the percentage of combined BTS and BTD kernels in a sample. The model had an R² of 0.64 and a standard error of prediction of 7.4%, showing it had some measurement ability for BTS and BTD. The SiLED‐SKS correctly classified and sorted out 90% of BTD and 66% of BTS for all 28 samples after three passes through the sorter. These instruments can serve as important tools for plant breeders and grading facilities of the wheat industry that require timely and objective determination and sorting of different levels of black tip present in wheat samples.     

Determining Weight and Moisture Properties of Sound and Fusarium‐Damaged Single Wheat Kernels by Near‐Infrared Spectroscopy

Single kernel moisture content (MC) is important in the measurement of other quality traits in single kernels because many traits are expressed on a dry weight basis. MC also affects viability, storage quality, and price. Also, if near‐infrared (NIR) spectroscopy is used to measure grain traits, the influence of water must be accounted for because water is a strong absorber throughout the NIR region. The feasibility of measurement of MC, fresh weight, dry weight, and water mass of single wheat kernels with or without Fusarium damage was investigated using two wheat cultivars with three visually selected classes of kernels with Fusarium damage and a range of MC. Calibration models were developed either from all kernel classes or from only undamaged kernels of one cultivar that were then validated using all spectra of the other cultivar. A calibration model developed for MC when using all kernels from the wheat cultivar Jagalene had a coefficient of determination (R²) of 0.77 and standard error of cross validation (SECV) of 1.03%. This model predicted the MC of the wheat cultivar 2137 with R² of 0.81 and a standard error of prediction (SEP) of 1.02% and RPD of 2.2. Calibration models developed using all kernels from both cultivars predicted MC, fresh weight, dry weight, or water mass in kernels better than models that used only undamaged kernels from both cultivars. Single kernel water mass was more accurately estimated using the actual fresh weight of kernels and MC predicted by calibrations that used all kernels or undamaged kernels. The necessity for evaluating and expressing constituent levels in single kernels on a mass/kernel basis rather than a percentage basis was elaborated. The need to overcome the effects of kernel size and water mass on single kernel spectra before using in calibration model development was also highlighted.     

Determining Vitreous Subclasses of Hard Red Spring Wheat Using Visible/Near‐Infrared Spectroscopy

The percentage of dark hard vitreous (DHV) kernels in hard red spring wheat is an important grading factor that is associated with protein content, kernel hardness, milling properties, and baking quality. The current visual method of determining DHV and non‐DHV (NDHV) wheat kernels is time‐consuming, tedious, and subject to large errors. The objective of this research was to classify DHV and NDHV wheat kernels, including kernels that were checked, cracked, sprouted, or bleached using visible/near‐infrared (Vis/NIR) spectroscopy. Spectra from single DHV and NDHV kernels were collected using a diode‐array NIR spectrometer. The dorsal and crease sides of the kernels were viewed. Three wavelength regions, 500–750 nm, 750–1,700 nm, and 500–1700 nm were compared. Spectra were analyzed by using partial least squares (PLS) regression. Results suggest that the major contributors to classifying DHV and NDHV kernels are light scattering, protein content, kernel hardness, starch content, and kernel color effects on the absorption spectrum. Bleached kernels were the most difficult to classify because of high lightness values. The sample set with bleached kernels yielded lower classification accuracies of 91.1–97.1% compared with 97.5–100% for the sample set without bleached kernels. More than 75% of misclassified kernels were bleached. For sample sets without bleached kernels, the classification models that included the dorsal side gave the highest classification accuracies (99.6–100%) for the testing sample set. Wavelengths in both the Vis/NIR regions or the NIR region alone yielded better classification accuracies than those in the visible region only.     

Protein Content of Bulk Wheat from Near‐Infrared Reflectance of Individual Kernels

Protein content of wheat by near‐infrared (NIR) reflectance of bulk samples is routinely practiced. New instrumentation that permits automated NIR analysis of individual kernels is now available, with the potential for rapid NIR‐based determinations of color, disease, and protein content, all on a single kernel (sk) basis. In the event that the protein content of the bulk sample is needed rather than that of the individual kernels, the present study examines the feasibility of estimating bulk sample protein from sk spectral readings. On the basis of 318 wheat samples of 10 kernels per sample, encompassing five U.S. wheat classes, the study demonstrates that with as few as 300 kernels bulk sample protein content may be estimated by sk NIR reflectance spectra at an accuracy equivalent to conventional bulk kernel NIR instrumentation.     

Moisture Effects on the Prediction Performance of a Single‐Kernel Near‐Infrared Deoxynivalenol Calibration

Fusarium head blight (FHB) is a serious disease in wheat that affects grain quality owing to the accumulation of mycotoxins such as deoxynivalenol (DON) in grains. Near‐infrared (NIR) spectroscopy has been used to develop techniques to estimate DON levels in single wheat kernels to facilitate rapid, nondestructive screening of FHB resistance in wheat breeding lines. The effect of moisture content (MC) variation on the accuracy of single‐kernel DON prediction by NIR spectroscopy was investigated. Sample MC considerably affected accuracy of the current NIR DON calibration by underestimating or overestimating DON at higher or lower moisture levels, respectively. DON in single kernels was most accurately estimated at 13–14% MC. Major NIR absorptions related to Fusarium damage were found around 1,198–1,200, 1,418–1,430, 1,698, and 1,896–1,914 nm. Major moisture related absorptions were observed around 1,162, 1,337, 1,405–1,408, 1,892–1,924, and 2,202 nm. Fusarium damage and moisture related absorptions overlapped in the 1,380–1,460 and 1,870–1,970 nm regions. These results show that absorption regions associated with water are often close to absorption regions associated with Fusarium damage. Thus, care must be taken to develop DON calibrations that are independent of grain MC.     

High‐Speed NIR Segregation of High‐ and Low‐Protein Single Wheat Seeds

Wheat breeders need a nondestructive method to rapidly sort high‐ or low‐protein single kernels from samples for their breeding programs. For this reason, a commercial color sorter equipped with near‐infrared filters was evaluated for its potential to sort high‐ and low‐protein single wheat kernels. Hard red winter and hard white wheat cultivars with protein content >12.5% (classed as high‐protein, 12% moisture basis) or < 11.5% (classed as low‐protein) were blended in proportions of 50:50 and 95:5 (or 5:95) mass. These wheat blends were sorted using five passes that removed 10% of the mass for each pass. The bulk protein content of accepted kernels (accepts) and rejected kernels (rejects) were measured for each pass. For 50:50 blends, the protein in the first‐pass rejects changed as much as 1%. For the accepts, each pass changed the protein content of accepts by ≈0.1%, depending on wheat blends. At most, two re‐sorts of accepts would be required to move 95:5 blends in the direction of the dominant protein content. The 95:5 and 50:50 blends approximate the low‐ and high‐protein mixture range of early generation wheat populations, and thus the sorter has potential to aid breeders in purifying samples for developing high‐ or low‐protein wheat. Results indicate that sorting was partly driven by color and vitreousness differences between high‐ and low‐protein fractions. Development of a new background specific for high‐ or low‐protein and fabrication of better optical filters for protein might help improve the sorter performance.     

Development of Nondestructive Screening Methods for Single Kernel Characterization of Wheat

The development of nondestructive screening methods for single seed protein, vitreousness, density, and hardness index has been studied for single kernels of European wheat. A single kernel procedure was applied involving, image analysis, near‐infrared transmittance (NIT) spectroscopy, laboratory density determination, single kernel characterization system (SKCS), and finally Kjeldahl protein determination on the crushed single kernels. Single kernel NIT spectroscopy showed excellent ability to determine protein content, and some ability for determination of single kernel vitreousness. Nondestructive determination of single kernel density, either based on NIT spectroscopy or based on image analysis and kernel weight, needs to be further improved for practical use. The use of SKCS hardness index as a true single kernel hardness reference in a NIT prediction model resulted in a poor predictability. However, by applying an averaging approach, in which single seed replicate measurements are mathematically simulated, a very good NIT prediction model was achieved. This suggests that the single seed NIT spectra contain hardness information, but that a single seed hardness method with higher accuracy is needed to achieve a good NIT prediction model for single kernel hardness.     

Selecting and Sorting Waxy Wheat Kernels Using Near‐Infrared Spectroscopy

An automated single kernel near‐infrared (NIR) sorting system was used to separate single wheat (Triticum aestivum L.) kernels with amylose‐free (waxy) starch from reduced‐amylose (partial waxy) or wild‐type wheat kernels. Waxy kernels of hexaploid wheat are null for the granule‐bound starch synthase alleles at all three Wx gene loci; partial waxy kernels have at least one null and one functional allele. Wild‐type kernels have three functional alleles. Our results demonstrate that automated single kernel NIR technology can be used to select waxy kernels from segregating breeding lines or to purify advanced breeding lines for the low‐amylose kernel trait. Calibrations based on either amylose content or the waxy trait performed similarly. Also, a calibration developed using the amylose content of waxy, partial waxy, and wild‐type durum (T. turgidum L. var durum) wheat enabled adequate sorting for hard red winter and hard red spring wheat with no modifications. Regression coefficients indicated that absorption by starch in the NIR region contributed to the classification models. Single kernel NIR technology offers significant benefits to breeding programs that are developing wheat with amylose‐free starches.     

Application of the Single‐Kernel Characterization System to Durum Wheat Testing and Quality Prediction

The Single Kernel Characterization System (SKCS 4100) measures single kernel weight, width, moisture content, and hardness in wheat grain with greater speed than existing methods and can be calibrated to predict flour starch damage and milling yield. The SKCS 4100 is potentially useful for testing applications in a durum improvement program. The mean SKCS 4100 kernel weight and moisture values from the analysis of 300 individual kernels gave good correlations with 1,000 kernel weight (r² = 0.956) and oven moisture (r² = 0.987), respectively. Although significant correlations were obtained between semolina mill yield and SKCS 4100 weight, diameter, and peak force, they were all very low and would be of little use for prediction purposes. Similarly, although there were significant correlations between some SKCS 4100 parameters and test weight and farinograph parameters, they too were small. The SKCS 4100 has been calibrated using either the single kernel hardness index or crush force profile to objectively measure the percentage vitreous grains in a sample with reasonable accuracy, and it correlates well with visual determination. The speed and accuracy of the test would be of interest to grain traders. An imprecise but potentially useful calibration was obtained for the prediction of semolina mill yield using the SKCS 4100 measurements on durum wheat. The SKCS 4100 is useful for some traits such as hardness, grain size and moisture for early‐generation (F3) selection in a durum improvement program.     

Single‐Kernel Near‐Infrared Analysis for Evaluating Wheat Samples for Fusarium Head Blight Resistance

This report describes a method to estimate the bulk deoxynivalenol (DON) content of wheat grain samples with the single‐kernel DON levels estimated by a single‐kernel near‐infrared (SKNIR) system combined with single‐kernel weights. The described method estimated the bulk DON levels in 90% of 160 grain samples to within 6.7 ppm of DON when compared with the DON content determined with the gas chromatography–mass spectrometry method. The single‐kernel DON analysis showed that the DON content among DON‐containing kernels (DCKs) varied considerably. The analysis of the distribution of DON levels among all kernels and among the DCKs of grain samples is helpful for the in‐depth evaluation of the effect of varieties or fungicides on Fusarium head blight (FHB) reactions. The SKNIR DON analysis and estimation of the single‐kernel DON distribution patterns demonstrated in this study may be helpful for wheat breeders to evaluate the FHB resistance of varieties in relation to their resistance to the spread of the disease and resistance to DON accumulation.     

近红外透反射光谱测定单粒稻种的蛋白质含量

为探索NIR光谱技术在水稻种子蛋白质含量分析中的应用,本研究细致分析了单粒稻种在不同光谱采集方式下的近红外光谱(NIRS)特征,并利用离子束诱变育种得到的水稻9311突变体库的种子,建立准确性较好的单粒糙米和单粒稻种的蛋白质定量模型。结果表明,与漫反射光谱采集方式下的单粒糙米蛋白质模型相比,透反射和透射光谱采集方式下能得到相关性较好的糙米蛋白质模型,其中单粒糙米蛋白质最优定量模型的决定系数(R²)为0.97,预测均方根误差(RMSEP)为0.27%。在单粒稻种中,由于种壳的反射作用,漫反射光谱采集方式下依然无法建立准确性高的蛋白质模型,透反射光谱采集方式下能够建立具有一定预测能力的蛋白质定量模型(RMSEP=0.81%),透射光谱采集方式下能够建立准确性高的蛋白质定量模型(R²=0.96,RMSEP=0.24%)。本研究结果为无损快速分析单粒稻种提供了一种解决方法。     

High‐Throughput Near‐Infrared Reflectance Spectroscopy for Predicting Quantitative and Qualitative Composition Phenotypes of Individual Maize Kernels

Near‐infrared reflectance (NIR) spectroscopy can be used for fast and reliable prediction of organic compounds in complex biological samples. We used a recently developed NIR spectroscopy instrument to predict starch, protein, oil, and weight of individual maize (Zea mays) seeds. The starch, protein, and oil calibrations have reliability equal or better to bulk grain NIR analyzers. We also show that the instrument can differentiate quantitative and qualitative seed composition mutants from normal siblings without a specific calibration for the constituent affected. The analyzer does not require a specific kernel orientation to predict composition or to differentiate mutants. The instrument collects a seed weight and a spectrum in 4–6 sec and can collect NIR data alone at a 20‐fold faster rate. The spectra are acquired while the kernel falls through a glass tube illuminated with broad spectrum light. These results show significant improvements over prior single‐kernel NIR systems, making this instrument a practical tool to collect quantitative seed phenotypes at high throughput. This technology has multiple applications for studying the genetic and physiological influences on seed traits.     

Discriminating Oat and Groat Kernels from Other Grains Using Near‐Infrared Spectroscopy

Oats and groats can be discriminated from other grains such as barley, wheat, rye, and triticale (nonoats) with near‐infrared spectroscopy. The two instruments tested herein were the manual version of the United States Department of Agriculture–Agricultural Research Service single‐kernel near‐infrared (SKNIR) instrument and the automated QualySense QSorter Explorer high‐speed sorter, both used in similar near‐infrared spectral ranges. Three linear discriminate self‐prediction models were developed: 1) oats versus groats + nonoats, 2) oats + groats versus nonoats, and 3) groats versus nonoats. For all three models, the SKNIR instrument showed high correct classification of oats or groats (94.5–100%), which was similar to results of the QSorter Explorer at 95.0–99.4%. The amount of nonoats that were misclassified as oats or groats was low for both instruments at 0–0.2% for the SKNIR instrument and 0.8–3.7% for the QSorter Explorer. Linear discriminate models from independent prediction and validation sets yielded classification accuracies of 91.6–99.3% (SKNIR) and 90.5–97.8% (QSorter Explorer). Small differences in classification accuracy were attributed to processing speeds between the two instruments: 3 kernels/s for the SKNIR instrument and 35 kernels/s for the QSorter Explorer. This indicated that both instruments are useful for quantifying grain sample compositions of oat and groat samples and that both could be useful tools for meeting consumer demand for gluten‐free or low‐gluten products. Discrimination between grains will help producers and manufacturers meet various regulatory requirements. Examples include requirements such as those from the U.S. Food and Drug Administration and the Commission of European Communities, in which gluten‐free oats or other products can only be labeled as nongluten if they contain gluten at less than 20 ppm, the established safe consumption limit for people with celiac disease. The QSorter Explorer is currently being used to meet these requirements.     

Near‐Infrared Spectroscopic Method for Identification of Fusarium Head Blight Damage and Prediction of Deoxynivalenol in Single Wheat Kernels

Fusarium Head Blight (FHB), or scab, can result in significant crop yield losses and contaminated grain in wheat (Triticum aestivum L.). Growing less susceptible cultivars is one of the most effective methods for managing FHB and for reducing deoxynivalenol (DON) levels in grain, but breeding programs lack a rapid and objective method for identifying the fungi and toxins. It is important to estimate proportions of sound kernels and Fusarium‐damaged kernels (FDK) in grain and to estimate DON levels of FDK to objectively assess the resistance of a cultivar. An automated single kernel near‐infrared (SKNIR) spectroscopic method for identification of FDK and for estimating DON levels was evaluated. The SKNIR system classified visually sound and FDK with an accuracy of 98.8 and 99.9%, respectively. The sound fraction had no or very little accumulation of DON. The FDK fraction was sorted into fractions with high or low DON content. The kernels identified as FDK by the SKNIR system had better correlation with other FHB assessment indices such as FHB severity, FHB incidence and kernels/g than visual FDK%. This technique can be successfully employed to nondestructively sort kernels with Fusarium damage and to estimate DON levels of those kernels. Single kernels could be predicted as having low (<60 ppm) or high (>60 ppm) DON with ≈96% accuracy. Single kernel DON levels of the high DON kernels could be estimated with R² = 0.87 and standard error of prediction (SEP) of 60.8 ppm. Because the method is nondestructive, seeds may be saved for generation advancement. The automated method is rapid (1 kernel/sec) and sorting grains into several fractions depending on DON levels will provide breeders with more information than techniques that deliver average DON levels from bulk seed samples.     

Predicting Wheat Quality Characteristics and Functionality Using Near‐Infrared Spectroscopy

The accuracy of using near‐infrared spectroscopy (NIRS) for predicting 186 grain, milling, flour, dough, and breadmaking quality parameters of 100 hard red winter (HRW) and 98 hard red spring (HRS) wheat and flour samples was evaluated. NIRS shows the potential for predicting protein content, moisture content, and flour color b* values with accuracies suitable for process control (R² > 0.97). Many other parameters were predicted with accuracies suitable for rough screening including test weight, average single kernel diameter and moisture content, SDS sedimentation volume, color a* values, total gluten content, mixograph, farinograph, and alveograph parameters, loaf volume, specific loaf volume, baking water absorption and mix time, gliadin and glutenin content, flour particle size, and the percentage of dark hard and vitreous kernels. Similar results were seen when analyzing data from either HRW or HRS wheat, and when predicting quality using spectra from either grain or flour. However, many attributes were correlated to protein content and this relationship influenced classification accuracies. When the influence of protein content was removed from the analyses, the only factors that could be predicted by NIRS with R² > 0.70 were moisture content, test weight, flour color, free lipids, flour particle size, and the percentage of dark hard and vitreous kernels. Thus, NIRS can be used to predict many grain quality and functionality traits, but mainly because of the high correlations of these traits to protein content.     

An Automated Near‐Infrared System for Selecting Individual Kernels Based on Specific Quality Characteristics

An automated sorting system was developed that nondestructively measured quality characteristics of individual kernels using near‐infrared (NIR) spectra. This single‐kernel NIR system was applied to sorting wheat (Triticum aestivum L.) kernels by protein content and hardness, and proso millet (Panicum miliaceum L.) into amylose‐bearing and amylose‐free fractions. Single wheat kernels with high protein content could be sorted from pure lines so that the high‐protein content portion was 3.1 percentage points higher than the portion with the low‐protein kernels. Likewise, single wheat kernels with specific hardness indices could be removed from pure lines such that the hardness index in the sorted samples was 29.4 hardness units higher than the soft kernels. The system was able to increase the waxy, or amylose‐free, millet kernels in segregating samples from 94% in the unsorted samples to 98% in the sorted samples. The portion of waxy millet kernels in segregating samples was increased from 32% in the unsorted samples to 55% after sorting. Thus, this technology can be used to enrich the desirable class within segregating populations in breeding programs, to increase the purity of heterogeneous advanced or released lines, or to measure the distribution of quality within samples during the marketing process.     

Differentiating Vitreous and Nonvitreous Durum Wheat Kernels by Using Near‐Infrared Spectroscopy

The vitreousnss of durum wheat is used by the wheat industry as an indicator of milling and cooking quality. The current visual method of determining vitreousness is subjective, and classification results between inspectors and countries vary widely. Thus, the use of near‐infrared (NIR) spectroscopy to objectively classify vitreous and nonvitreous single kernels was investigated. Results showed that classification of obviously vitreous or nonvitreous kernels by the NIR procedure agreed almost perfectly with inspector classifications. However, when difficult‐to‐classify vitreous and nonvitreous kernels were included in the analysis, the NIR procedure agreed with inspectors on only 75% of kernels. While the classification of difficult kernels by NIR spectroscopy did not match well with inspector classifications, this NIR procedure quantifies vitreousness and thus may provide an objective classification means that could reduce inspector‐to‐inspector variability. Classifications appear to be due, at least in part, to scattering effects and to starch and protein differences between vitreous and nonvitreous kernels.     

Near‐Infrared Hyperspectral Imaging of Fusarium‐Damaged Oats (Avena sativa L.)

The feasibility of hyperspectral imaging (HSI) to detect deoxynivalenol (DON) content and Fusarium damage in single oat kernels was investigated. Hyperspectral images of oat kernels from a Fusarium‐inoculated nursery were used after visual classification as asymptomatic, mildly damaged, and severely damaged. Uninoculated kernels were included as controls. The average spectrum from each kernel was paired with the reference DON value for the same kernel, and a calibration model was fitted by partial least squares regression (PLSR). To correct for the skewed distribution of DON values and avoid nonlinearities in the model, the DON values were transformed as DON* = [log(DON)]³. The model was optimized by cross‐validation, and its prediction performance was validated by predicting DON* values for a separate set of validation kernels. The PLSR model and linear discriminant analysis classification were further used on single‐pixel spectra to investigate the spatial distribution of infection in the kernels. There were clear differences between the kernel classes. The first component separated the uninoculated and asymptomatic from the severely damaged kernels. Infected kernels showed higher intensities at 1,925, 2,070, and 2,140 nm, whereas noninfected kernels were dominated by signals at 1,400, 1,626, and 1,850 nm. The DON* values of the validation kernels were estimated by using their average spectra, and the correlation (R) between predicted and measured DON* was 0.8. Our results show that HSI has great potential in detecting Fusarium damage and predicting DON in oats, but it needs more work to develop a model for routine application.     

Estimation of the Deoxynivalenol and Moisture Contents of Bulk Wheat Grain Samples by FT‐NIR Spectroscopy

Deoxynivalenol (DON) levels in harvested grain samples are used to evaluate the Fusarium head blight (FHB) resistance of wheat cultivars and breeding lines. Fourier transform near‐infrared (FT‐NIR) calibrations were developed to estimate the DON level and moisture content (MC) of bulk wheat grain samples harvested from FHB screening trials. Grains in a rotating glass petri dish were scanned in the 10,000–4,000 cm⁻¹ (1,000–2,500 nm) spectral range using a Perkin Elmer Spectrum 400 FT‐IR/FT‐NIR spectrometer. The DON calibration predicted the DON levels in test samples with R ² = 0.62 and root mean square error of prediction (RMSEP) = 8.01 ppm. When 5–25 ppm of DON was used as the cut‐off to classify samples into low‐ and high‐DON groups, 60.8–82.3% of the low‐DON samples were correctly classified, whereas the classification accuracy of the high‐DON group was 82.3–94.0%. The MC calibration predicted the MC in grain samples with R ² = 0.98 and RMSEP = 0.19%. Therefore, these FT‐NIR calibrations can be used to rapidly prescreen wheat lines to identify low‐DON lines for further evaluation using standard laboratory methods, thereby reducing the time and costs of analyzing samples from FHB screening trials.